Non-invasive stem cell tracking in hindlimb ischemia animal model using bio-orthogonal copper-free click chemistry
- Non-invasive stem cell tracking in hindlimb ischemia animal model using bio-orthogonal copper-free click chemistry
- 김광명; 이상민; 이장욱; 이시연; 윤화인; 이지영; 박순정; 구희범; 문성환; 이혁진; 조용우; 강선웅; 이상엽
- Cell labeling and tracking; Bio-orthogonal copper-free click chemistry; Metabolic glycoengineering; Mesenchymal stem cell; Hindlimb ischemia
- Issue Date
- Biochemical and biophysical research communications
- VOL 479, NO 4-786
- Labeling of stem cells aims to distinguish transplanted cells from host cells, understand in vivo fate of transplanted cells, particularly important in stem cell therapy. Adipose-derived mesenchymal stem cells (ASCs) are considered as an emerging therapeutic option for tissue regeneration, but much remains to be understood regarding the in vivo evidence. In this study, a simple and efficient cell labeling method for labeling and tracking of stem cells was developed based on bio-orthogonal copper-free click chemistry, and it was applied in a mouse hindlimb ischemia model. The human ASCs were treated with tetra-acetylated N-azidoacetyl-d-mannosamine (Ac4ManNAz) to generate glycoprotein with unnatural azide groups on the cell surface, and the generated azide groups were fluorescently labeled by specific binding of dibenzylcyclooctyne-conjugated Cy5 (DBCO-Cy5). The safe and long-term labeling of the hASCs by this method was first investigated in vitro. Then the DBCO-Cy5-hASCs were transplanted into the hindlimb ischemia mice model, and we could monitor and track in vivo fate of the cells using optical imaging system. We could clearly observe the migration potent of the hASCs toward the ischemic lesion. This approach to design and tailor new method for labeling of stem cells may be useful to provide better understanding on the therapeutic effects of transplanted stem cells into the target diseases.
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